Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2003-10-31 to 2003-10-31
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Only a study summary was available for review which provided limited details on the test substance and methodology; however, sufficient information was provided to deem the study reliable with restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Principles of method if other than guideline:
The ocular irritancy potential of the test material was assessed using the Rabbit Enucleated Eye Test (REET). This method involved the apploication of the test material onto the cornea of the enucleated eye.
Direct effect on the cornea is assessed by evaluation of corneal thickness, opacity, alteration of corneal epithelium, and fluorescein uptake. The data for all endpoints are assessed and an estimate of ocular irritancy potential is made.
GLP compliance:
no
Remarks:
performed in facility operating to GLP, but no formal claim or audit by the QA unit

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study reports): T000836 (JNJ-16250351-AAH)
- Physical state: solid
- Appearance: white powder
Specific details on test material used for the study:
no data

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Five enucleated eyes, obtained from the New Zealand White strain of rabbit.
- The eyes were maintained at a temeprature of 32°C +/-1.5°C within the superfusion apparatus, except for 180 and 240 minutes post dosing when the temperature was slightly below the stated range. This deviation from the Standard Test Method was considered not to affect the purpose or integrity of the study.

Test system

Vehicle:
not specified
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approximately 65 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): no data
- Concentration (if solution): 0.9% sodium chloride
Duration of treatment / exposure:
single application
Number of animals or in vitro replicates:
3 treated eyes, 2 control eyes
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no data

OBSERVATION TIME POINTS
- corneal opacity and corneal epithelium condition: 60, 120, 180 and 240 min
- fluorescein uptake: 240 min
- corneal swelling: 60, 120 and 240 min

SCORING SYSTEM:
- Method for Evaluation of Ocular Irritation by Slit-Lamp Biomicroscopic Examination (McDonald Shadduck Score System)
- The scoring scheme measures the severity of corneal cloudiness and the area of the cornea involve d. Severity of corneal cloudiness is graded as follows:
0 = Normal cornea. Appears with the slit-lamp as having a bright grey line on the epithelial surface and a bright grey appearance of the stroma.
1 = Some loss of transparency. Only the anterior half of the stroma is involved as observed with an optical section of the slit-lamp. The underlying structures are clearly visible with diffuse illumination, although some cloudiness can be readily apparent with diffuse illumination.
2 = Moderate loss of transparency. In addition to involving the anterior stroma, the cloudiness extends all the way to the endothelium. The stroma has lost its marble-like appearance and is homogeneously white. With diffuse, illumination, underlying structures are clearly visible.
3 = Involvement of the entire thickness of the stroma. With optical section, the endothelial surface is still visible. However, with diffuse illumination the underlying structures are just visible.
4 = Involvement of the entire thickness of the stroma. With the optical section cannot clearly visualise the endothelium. With diffuse illumination, the underlying structures cannot be seen.
5
The surface of the cornea relative to the area of cloudiness is divided into five grades from 0 to 4:
0 = Normal cornea with no area of cloudiness
1 = 1 to 25% area of stromal cloudiness
2 = 26 to 50% area of stromal cloudiness
3 = 51 to 75% area of stromal cloudiness
4 = 76 to 100% area of stromal cloudiness
FLUORESCEIN
- The use of fluorescein is a valuable aid in defining epithelial damage for fluorescein staining. The a rea can be judged as a 0 to 4 scale using the same terminology as for corneal cloudiness. The intensity of fluorescein staining can be divided into a 0 to 4 scale:
0 = Absence of fluorescein staining
1 = Slight fluorescein staining confined to a small focus. With diffuse illumination the underlying struc tures are clearly visible, although there is some loss of detail.
2 = Moderate fluorescein staining confined to a small focus. With diffuse illumination the underlying str uctures are clearly visible, although there is some loss of detail.
3 = Marked fluorescein staining. Staining may involve a larger portion of the cornea. With diffuse illu mination underlying structures are barely visible but are not completely obliterated.
4 = Extreme fluorescein staining. With diffuse illumination the underlying structures cannot be seen.

TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - cloudy
Run / experiment:
60 minutes post dosing
Value:
0
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - area
Run / experiment:
60 minutes post dosing
Value:
0
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - cloudy
Run / experiment:
120 minutes post dosing
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - area
Run / experiment:
120 minutes post dosing
Value:
3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - cloudy
Run / experiment:
180 post dosing
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - area
Run / experiment:
180 minutes post dosing
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of 3 eyes - cloudy
Run / experiment:
240 minutes post dosing
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
cornea opacity score
Remarks:
mean of teyes - area
Run / experiment:
240 minutes post dosing
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
fluorescein retention score
Remarks:
mean of 3 eyes (intensity)
Run / experiment:
240 minutes post dosing
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
fluorescein retention score
Remarks:
mean of 3 eyes (area)
Run / experiment:
240 minutes post dosing
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
percent corneal swelling
Remarks:
mean of 3 eyes
Run / experiment:
60 minutes post dosing
Value:
14.9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
percent corneal swelling
Remarks:
mean of 3 eyes
Run / experiment:
120 minutes post dosing
Value:
19.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
percent corneal swelling
Remarks:
mean of 3 eyes
Run / experiment:
240 minutes post dosing
Value:
29.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Other effects / acceptance of results:
negative control eyes:
- corneal opacity: 0 (mean of 2 eyes)
- corneal epithelium condition: normal (2 eyes)
- fluorescein uptake: 0 (mean of 2 eyes)
- corneal swelling: 4.7 at 60 min post dosing, 4.4 at 120 minutes post dosing and 2.8 at 240 minutes post dosing

Corneal epithelium condition of test item treated eyes was normal at all timepoints

In vivo

Irritant / corrosive response data:
.

Any other information on results incl. tables

The test item adhered to the surface of the cornea.  

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
The test material was considered to have the potential to cause severe ocular irritancy in vivo.